Cloning large DNA fragments using the bacterial artificial chromosome (BAC) library system has revolutionized and accelerated our ability to physically map, positionally clone and sequence whole genomes. Due to the difficulty in cloning large DNA fragments, only a handful of laboratories have been able to produce deep-coverage large insert libraries on a routine basis. This bottleneck has resulted in a limited number of BAC libraries that are available to the research community that can be used for gene discovery, survey sequencing and comparative genome studies. To increase the world?s BAC library construction capacity and reduce costs, the NIH has posted a request for applications (RFA.HG-Ol-002) entitled "BAC Library Production" which is the subject of this application. The specific aims of our thee-year proposal are: BAC Library Production: Develop BAC library construction capacity to construct 36 human genome size ( about3000 Mb) 10X genome equivalent BAC libraries. In year 1, we will produce 6 libraries at $0.50/clone, in year 2, 12 libraries will be produced at $0.375/clone and in year three 18 libraries will he produced at $0.25/clone. The initial average insert size will hover between 130 - 150 kb and we will strive to increase our average insert sizes to 170 - 200 kb in subsequent years. Libraries, filters and clones will be distributed through our CUGI BAC/EST Resource Center on a cost recovery basis (www.genome.clemson.edu). BAC Library Research and Development: We will systematically develop technologies to optimize and streamline BAC library construction procedures in order to reduce the cost of making libraries while maintaining quality and insert size. Also, technologies will be developed to routinely achieve insert sizes of 160kb or greater. In addition, a robust tool kit will be established to produce high-quality megabase-size DNA from animals. This will include the development of a "BIG DNA To-Go" kit to produce high-quality megabase-size DNA on location where the lab needing a library will provide the tissue(s) required to make the library. Bioinformatics Support: A laboratory information management/bar coding system will be developed to aid in tracking library production and distribution and a robust online BAC library ordering system and database will also be developed.